High-efficiency transformation of bacterial cells by electroporation.

Abstract:

:We have developed a method for efficiently generating transient pores in the outer membranes of Escherichia coli K-12 derivatives by using a new type of electroporation apparatus. The pores are large enough and persist long enough to facilitate the equilibration of plasmid molecules between the intracellular and extracellular spaces. The method has been used to transform bacterial cells with an efficiency greater than 10(9) transformants per microgram of plasmid. It has also been used to extract intact plasmid from transformed cells with efficiencies comparable to those of the traditional alkaline lysis or CsCl equilibrium density gradient techniques. The technique is simple and rapid, allowing a transformation or the preparation of microgram quantities of plasmid to be accomplished in minutes.

journal_name

J Bacteriol

journal_title

Journal of bacteriology

authors

Calvin NM,Hanawalt PC

doi

10.1128/jb.170.6.2796-2801.1988

subject

Has Abstract

pub_date

1988-06-01 00:00:00

pages

2796-801

issue

6

eissn

0021-9193

issn

1098-5530

journal_volume

170

pub_type

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